作　　者： ; ; ;

机构地区： 南方医科大学南方医院

出　　处： 《南方医科大学学报》 2007年第2期205-207,共3页

摘　　要： 目的构建人表皮生长因子(EGF)-linker-天花粉蛋白(TCS)融合蛋白的表达载体,在大肠杆菌中表达并纯化EGF-linker-TCS融合蛋白。方法用PCR方法扩增EGF及柔性连接肽Linker基因片段,插入质粒PQE30-TCS中,转化至大肠杆菌M15中,表达该融合蛋白(EGF-linker-TCS)。表达产物经Ni-NTAAgrose亲和层析柱纯化。结果PQE30-EGF-linker-TCS蛋白在大肠杆菌M15中获得稳定、高效的融合表达。目的蛋白占细胞总蛋白的40%左右,以可溶性形式存在,表达上清柱纯化后纯度达95%以上。结论应用基因工程技术,成功地构建了融合蛋白EGF-linker-TCS的表达载体,表达并纯化了该融合蛋白,为进一步研究其结构功能关系和肿瘤临床应用奠定了基础。 Objective To construct a recombinant expression vector of the fusion protein epidermal growth factor （EGF）- Linker-trichosanthin （TCS） and achieve its expression in E.coli to obtain purified EGF-Iinker-TCS fusion protein. Methods The gene fragments of EGF-linker were amplified by PCR and inserted into the expression plasmid PQE30-TCS, followed by transformation of the recombinant plasmid into E.coli MI 5 for expression of the fusion protein. Ni-FF column chromatography was utilized for purification of the expressed product. Results The recombinant plasmid PQE30-EGF- linker-TCS was stably and highly expressed in E. coli MI5. The expressed product existed in the form of soluble protein accounting for about 40% of total cellular protein and reached a purity of above 95% after purification with Ni-FF column chromatography. Conclusion The recombinant plasmid PQE30/EGF-Iinker-TCS has been successfully constructed, which provides a basis for further structural and functional study of EGF and TCS and their potential clinical application for cancer therapy.

关 键 词： 表皮生长因子 天花粉蛋白 重组融合蛋白 纯化

领　　域： [生物学]